Sanger测序法和Snapshot法检测乳腺癌BIM缺失多态性比较分析  

Study on The Concordance of BIM Deletion Polymorphism Tests by Sanger Sequencing and Snapshot Methods in Breast Cancer

作　　者：梁红玲 李洪胜 黄健清[2] 苏健[3] 金田恩 韩国栋 敖翔 邱霓 谭小军 LIANG Hong-ling;LI Hong-sheng;HUANG Jian-qing;SU Jian;JIN Tian-en;HAN Guo-dong;AO Xiang;QIU Ni;TAN Xiao-jun(Department of Breast Surgery,Affiliated Cancer Hospital&Institute of Guangzhou Medical University,Guangzhou 510095,China;Guangzhou Medical University,Guangzhou 510006,China;Medical Research Center,Guangdong General Hospital,Guangdong Academy of Medical Sciences,Guangdong Lung Cancer Institute,Guangzhou 510080,China;Guangzhou DAAN Clinical Laboratory Center,Guangzhou 510520,China)

机构地区：[1]广州医科大学附属肿瘤医院乳腺外科,广州510095 [2]广州医科大学,广州510006 [3]广东省人民医院医学研究部广东省医学科学院广东省肺癌研究所,广州510080 [4]广州达安临床检验中心,广州510520

出　　处：《循证医学》2018年第6期370-375,共6页The Journal of Evidence-Based Medicine

基　　金：广东省医学科学技术研究基金项目指令性课题资助项目(C2015039);广州市西医类一般引导资助项目(20151A011088)

摘　　要：目的比较分析Sanger测序法和Snapshot法检测乳腺癌BIM缺失多态性的一致性。方法共收集336例乳腺癌患者手术标本制备成石蜡包埋的蜡块,提取DNA后采用Sanger测序法和Snapshot法检测乳腺癌患者中BIM基因缺失多态性突变率。采用卡方检验分析BIM缺失多态性在不同临床特征患者中的比例情况。采用配对卡方检验和κ系数检验分析两种方法的一致性。结果 Sanger测序法检测出39例患者BIM基因缺失,阳性率为11.61%;Snapshot法检测出47例患者BIM基因缺失,阳性率为13.99%。Sanger测序法检测出的BIM突变状态在不同年龄阶段(χ~2=6.242,P=0.044)和孕激素表达水平(χ~2=4.070,P=0.044)的乳腺癌中差异有统计学意义。配对卡方检验显示两种检测方法差异无统计学意义(P=0.152),Kappa检验结果表明两种方法的一致性较好(吻合系数κ=0.695,P=0.000)。结论在中国乳腺癌患者中BIM缺失多态性的比例约为11.61%。Sanger测序法和Snapshot法检测乳腺癌BIM缺失多态性具有较好的一致性,可以相互补充。Objective The role of this essay is to explore the cocordance of Sanger sequencing and Snapshot methods for detecting the deletion polymorphism of BIM gene in breast cancer.Methods Surgical cancer tissue samples of a total of 336 cases with breast cancer were collected and preserved as formalin fixed paraffin embedded wax blocks.Genomic DNA was extracted and BIM gene deletion polymorphic in these breast cancer patients were detected by Sanger sequencing and Snapshot methods.Chi?square test with cross?tabular data was used to analyze the proportion of BIM deletion polymorphisms in patients with different clinical characteristics.Chi square test and Kappa coefficient test were used to detect the concordance degree of the two methods.Results In these 336 cases of breast cancers,11.61%(39/336)cases were tested as positive with BIM deletion by Sanger sequencing.13.99%(47/336)patients were positive with BIM deletion porlymorphism by Snapshot method.The Sanger sequencing method detected statistically significant differences in the expression of BIM mutations in breast cancer at different ages(c2=6.242,P=0.044)and progestrone receptor levels(c2=4.070,P=0.044).The McNemar test showed the two methods had no statistical difference in the positivity of BIM gene deletion polymorphism(P=0.152).Kappa coefficient test also indicated concordance between the two methods(k=0.695,P=0.000).Conclusions In this Chinese cohort of breast cancers,the BIM gene deletion polymorphism is around 11.61%.Sanger sequencing and Snapshot methods had consistency and might be complementary in the detection of BIM deletion polymorphism in breast cancer.

关 键 词：乳腺癌 BIM基因 Sanger测序法 Snapshot检测法 

分 类 号：R737.9[医药卫生—肿瘤]